• The Korean Journal of Physiology and Pharmacology
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• v.24 no.1
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• pp.101-110
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• 2020

Transient receptor potential canonical 4 (TRPC4) channel is a nonselective calcium-permeable cation channels. In intestinal smooth muscle cells, TRPC4 currents contribute more than 80% to muscarinic cationic current (mIcat). With its inward-rectifying current-voltage relationship and high calcium permeability, TRPC4 channels permit calcium influx once the channel is opened by muscarinic receptor stimulation. Polyamines are known to inhibit nonselective cation channels that mediate the generation of mIcat. Moreover, it is reported that TRPC4 channels are blocked by the intracellular spermine through electrostatic interaction with glutamate residues (E728, E729). Here, we investigated the correlation between the magnitude of channel inactivation by spermine and the magnitude of channel conductance. We also found additional spermine binding sites in TRPC4. We evaluated channel activity with electrophysiological recordings and revalidated structural significance based on Cryo-EM structure, which was resolved recently. We found that there is no correlation between magnitude of inhibitory action of spermine and magnitude of maximum current of the channel. In intracellular region, TRPC4 attracts spermine at channel periphery by reducing access resistance, and acidic residues contribute to blocking action of intracellular spermine; channel periphery, E649; cytosolic space, D629, D649, and E687.

• The Korea Journal of Herbology
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• v.22 no.4
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• pp.65-73
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• 2007

Objective : Salvia miltiorrhiza Bunge (Labiatae) (SM), an eminent herbal plant, has been widely used in traditional Chinese medicine for the treatment of vascular diseases such as hypertension. The aim of this study was to determine whether SM inhibits production of nitrite, an index of NO, and proinflammatory cytokines in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages. And this study investigated whether or not SM could reduce tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$)-induced inflammatory response in human vascular aortic smooth muscle cells (HASMC) and umbilical vein endothelial cells (HUVEC). Methods : Cytotoxic activity of SM on RAW 264.7 cells was using 5-(3-caroboxymeth-oxy phenyJ)-2H-tetra-zolium inner salt (MTS) assay. We measured the NO production using Griess Reagent System. Production of Proliflammatory cytokines was measured by Enzyme-Linked Immunosorbent Assay (ELISA). Results : Our results indicated that SM significantly inhibited the LPS-induced NO production accompanied by an attenuation of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), IL-6 and monocyte chemoattractant protein (MCP)-1 formation in macrophages. SM decreased TNF-${\alpha}$-induced IL-8, IL-6 production, and intracellular adhesion molecule (ICAM)-1 and vascular cell adhesion molecule (VCAM)-1 expression. Conclusion : These results indicate that SM has potential as an anti-inflammatory agent.

• Journal of the Korean Society of Food Science and Nutrition
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• v.23 no.3
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• pp.530-539
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• 1994

All lipoproteins are made up of three major classes of lipids : triglycerides, cholesterol, and phospholipids. Lipoproteins vary in their relative content of these lipids as well as in size and protein content. Human low density lipoprotein (LDL) is a main carrier for cholesterol in the blood stream, and it is well established that cholesterol deposits in the arteries stem primarily from LDL and that increased levels of plasma LDL correlated with in increased risk of atherosclerosis. Various lines of research provide strong evidence that lDL may become oxidized in vivo and that oxidized-LDL is the species involved in the formation of early atherosclerotic lesions. the most crucial findings in this context are the following : (1) Oxidized -LDL has chemotactic properties and if present in the intimal space of the arteries would recruit blood monocytes which then can develop into tissue macrophages ; (2) marcrophages take up oxidized-LDL unregulated to from lipid laden foam cells ; (3) Oxdized-LDLis highly cytotoxic and could be responsible for damage of the endothelial layer and for the destruction of smooth muscle cells.

• Korean Journal of Clinical Laboratory Science
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• v.40 no.2
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• pp.118-128
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• 2008

The aim of this study was to investigate the effect of electrical stimulation on healing of impaired wound and alteration of mast cells in experimental diabetic rats. Thirty male Sprague-Dawley rats were divided into three groups : incision (control), diabetes+incision (diabetes) and diabetes + incision + electrical stimulation (D/ES). Diabetes was induced in rats by streptozotocin (STZ) injection (60 mg/kg, one time) and 20 mm length incision wounds were created on the back after shaving hair. The electrical stimulation rats were treated with a current intensity of 30~50 V at 120 pps and $140{\mu}s$ for 10 days from 3 days after STZ injection. The lesion and adjacent skin tissues were fixed with 10% buffered formalin, embedded with paraffin. For wound healing analysis, hematoxylin-eosin (HE) and picrosirius red staining were performed. Mast cells (MC) were stained with toluidine blue (pH 0.5) and quantified at ${\times}200$ using a light microscope. The density of keratinocyte proliferation and microvessels in skin tissues were analyzed using a computerized image analysis system on sections immunostained with proliferative cell nuclear antigen (PCNA) and ${\alpha}$-smooth muscle actin (${\alpha}$-SMA), respectively. The results showed that the wound healing rate, collagen density and neoepidermis thickness, density of PCNA-positive cells and density of ${\alpha}$-SMA-positive vessels were significantly higher in D/ES rats than in diabetic rats. The density of MCs and degranulated MCs in D/ES rats were also significantly higher than those in diabetic rats. These findings suggest that the electrical stimulation may promote the tissue repair process by accelerating collagen production, keratinocyte proliferation and angiogenesis in the diabetic rats, and MCs are required for wound healing of skin in rats.

• BMB Reports
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• v.30 no.2
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• pp.106-112
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• 1997

The effect of cryopreservation on extracellular matrix was studied with the ultimate objective of permiting a prediction of the tendency of aorta conduit tissue to calcify following transplantation. Cryopreserved and fresh porcine aorta conduit tissues were extracted using guanidine-hydrochloride (Gdn-HCl) followed by sequential digestion of the tissues with collagenase, elastase, and papain. Glycosaminoglycans (GAGs) of the proteoglycans (PGs) were isolated and quantitated. Gdn-HCl extracted about 61% and 62% of the total GAG (proteoqlycan) material from cryopreserved and fresh tissues, respectively. Collagenasesolubilized proteoglycans from Gdn-HCl extracted tissue represented 20% and 13%, respectively, of the total GAGs present in cryopreserved and fresh tissues. Subsequent elastase hydrolysis of collagenase-digested tissue released about 11% of total GAGs from cryopreserved tissue and 16% from fresh tissue. The remaining 8%, from cryopreserved tissue, and 9%, from fresh tissue, of the total GAGs were obtained after using a papain hydrolysis. There was essentially no difference between fresh and cryopreserved tissues in the relative distribution of proteoglycans in the extracts and digestions except in the initial digestion step where more proteoglycans were obtained from collagenase solubilization of cryopreserved tissue than fresh tissue (p<0.05). The histologic status of the fresh and cryopreserved porcine aortic conduit did not differ markedly. The normal tissue architecture was not affected markedly by the cryopreservation procedure as neither alteration of elastic structure, fibrous proteins nor alteration of nuclear distribution or smooth muscle cell morphology was detected. Quantitative tissue mineral studies revealed that the mean calcium content of the cryopreserved aorta conduit tissue $(165{\pm}3\;{\mu}g/g\;wet\;tissue)$ was higher than that of the fresh tissue $(105{\pm}4\;{\mu}g/g\;wet\;tissue)$ $(p<0.05)$. The mean phosphorus content was $703{\pm}35\;{\mu}g$ wet tissue from cryopreserved tissue and $720{\pm}26\;{\mu}g$ wet tissue from fresh tissue. The study indicates that there is no significant alteration in the distribution of PGs in properly cryopreserved tissue, but the total calcium level appears to be increased in tissue cryopreserved by the cryopreservation process used in this study.

• The Korean Journal of Physiology and Pharmacology
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• v.17 no.5
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• pp.463-468
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• 2013

Acute hypoxia induces contraction of pulmonary artery (PA) to protect ventilation/perfusion mismatch in lungs. As for the cellular mechanism of hypoxic pulmonary vasoconstriction (HPV), hypoxic inhibition of voltage-gated $K^+$ channel (Kv) in PA smooth muscle cell (PASMC) has been suggested. In addition, our recent study showed that thromboxane $A_2$ ($TXA_2$) and hypoxia-activated nonselective cation channel ($I_{NSC}$) is also essential for HPV. However, it is not well understood whether HPV is maintained in the animals exposed to ambient hypoxia for two days (2d-H). Specifically, the associated electrophysiological changes in PASMCs have not been studied. Here we investigate the effects of 2d-H on HPV in isolated ventilated/perfused lungs (V/P lungs) from rats. HPV was almost abolished without structural remodeling of PA in 2d-H rats, and the lost HPV was not recovered by Kv inhibitor, 4-aminopyridine. Patch clamp study showed that the hypoxic inhibition of Kv current in PASMC was similar between 2d-H and control. In contrast, hypoxia and $TXA_2$-activated $I_{NSC}$ was not observed in PASMCs of 2d-H. From above results, it is suggested that the decreased $I_{NSC}$ might be the primary functional cause of HPV disappearance in the relatively early period (2 d) of hypoxia.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.11
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• pp.4977-4982
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• 2016

Introduction: Elevated serum interleukin (IL) 6 has been reported in patients infected with the hepatitis C virus (HCV), but it remains debatable whether this influences the production of autoantibodies and the biochemical profile of HCV disease. Therefore, this current study was conducted to evaluate the relationship between IL-6 and circulating autoantibody levels in HCV positive patients. Methods: Levels of IL-6 in serum samples from 102 patients with HCV and 103 normal controls were determined by enzyme linked immunosorbent assay (ELISA). Autoantibodies were detected by immunofluorescence. Results: Levels of IL-6 were significantly higher (p=0.028) in patients infected with (HCV) compared with normal group. Autoantibodies were noted in in 43.1% of the patients; of these, 23.5% featured anti-nuclear antibodies (ANA+), 16.7% anti-smooth muscle antibodies (ASMA+), 7.8% anti-mitochondrial antibodies (AMA+), 17.6% anti-parietal cell antibodies (APCA+), 7.8% anti canalicular antibodies, and 2.9% anti reticulin antibodies (ARA+). No patients were found to be positive for anti-brush border antibodies (ABBA) or anti-ribosomal antibodies. (ARiA). No links with IL-6 levels were apparent. Conclusions: IL-6 levels are increased in patients infected with HCV disease and could influence the production of autoantibodies. However, this study did not provide evidence of a specific relationship between IL6 and circulating autoantibodies in such cases.

• The Korean Journal of Physiology and Pharmacology
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• v.10 no.3
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• pp.161-165
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• 2006

NO and cyclooxygenase-2 (COX-2) are contributes to vascular inflammation induced by various stimulation. The mechanism, which explains a linkage between NO and COX-2, could be of importance in promoting pathophysiological conditions of vessel. We investigated the effects of NO donors on the COX-l and COX-2 mRNA/protein expression, as well as the nitrite production in culture medium of vascular smooth muscle cell (VSMC). VSMC was primarily cultured from thoracic aorta of rat. In this experiments, COX-l and COX-2 mRNA/protein expressions were analysed and nitrite productions were investigated using Griess reagent. VSMC did not express COX-2 protein in basal condition (Nonlipopolysaccharide (LPS) stimulated). In LPS-stimulated experiments, after 3 hours of NO donor pretreatment, LPS $10{\mu}g/ml$ was treated for 24 hours. COX-l protein expressions were unchanged by SNP and NOR-3. NOR-3 significantly increased COX-2 mRNA/protein expression under LPS stimulation. In contrast, SNP did not increase COX-2 mRNA/protein expression under LPS stimulation. Nitrite production was higher in NOR-3 treatment than SNP treatment under LPS stimulation. These results suggest that the expression of COX-2 in VSMC is regulated by NOR-3, COX-2 expressions were depending on the types of NO donor and LPS stimulation in VSMC.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.16 no.4
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• pp.397-418
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• 1994

Microvascular surgery has been widely used in the clinical field of replantation and reconstructive surgery. Since the last 20 years, microsurgical techniques and instruments have been rapidly developed and the success rate is remarkably increased. But thrombotic occlusion of vessels remains the major reason for clinical failure. The change of vessel wall is the most important factor in thrombus formation. If we can reduce the traumatic changes in the vessel walls during surgery, the success rate can be markedly increased. For this study, femoral arteries and veins of 36 Sprague-Dawley rats with average weights of 300gm were used. The author observed the histological changes and healing process in the anastomostic site after 1 hour, 24 hours, 1, 2, 3 and 4 weeks under light microscopy and scanning electron microscopy. The results were as follows : 1. The patency rate was 100% in femoral arteries and 85% in femoral vein. 2. At the early stages after microvascular anastomosis, the loss of endothelial cell in the vessel walls was observed in the wide area including anastomotic site. In scanning electron microscopic finding the anastomotic site was covered with much fibrin, many red blood cells and some platelets. 3. At 1st week, new endothelial cells were formed toward anastomotic site and at 3rd week, the anastomotic site was completely covered by new endothelial cells. At 4th week, the complete endothelialization over the threads was observed. 4. The media extended from the anastomotic site toward the end of the specimen. At later stages, the extent of media necrosis was markedly decreased. But the media necrosis of anastomotic site was not regenerated till 4th week. 5. Intimal hyperplasia appeared at 1st week and increased till 4th week. The layer consisted of endothelialization the most luminal layers and smooth muscle in the deeper layers. But in veins, the response was less pronounced than in arteries. 6. Foreign body granuloma remained during 4 weeks and aneurysm was observed at 3rd week in artery. In aneurismal wall, media necrosis, loss of elastic lamina and intimal hyperplasia were seen.

• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.3
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• pp.305-310
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• 2007

This study was designed to investigate the effects of KH-305 on erectile dysfunction in young rats, via nitric oxide (NO)-cGMP pathways. After oral administration of the KH-305 mixture (50, 100, 200, 300 mg/kg) to young rats for 10 days, NOS and SOD protein expressions in penile tissue and testosterone in plasma were measured. cGMP degradation was also investigated using bovine vascular smooth muscle cells pretreated with an NO donor, S-nitroso-N-Acetylpenicillamine (SNAP). The penile expression levels of nNOS and eNOS-dependent NOS activities as well as SOD preventing oxidative stress by overproduction of NO were increased significantly. Also, the concentration of testosterone in the plasma was increased. In vitro, cGMP concen-trations were decreased dose dependently in the KH-305. These results suggest that KH-305 may be useful in erectile dysfunction.